Novel use of a chemically modified siRNA for robust and sustainable in vivo gene silencing in the retina

Abstract Despite efficient and specific in vitro knockdown, more reliable and convenient methods for in vivo knockdown of target genes remain to be developed particularly for retinal research. Using commercially available and chemically modified siRNA so-called Accell siRNA, we established a novel i...

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Autores principales:	Takazumi Taniguchi, Ken-ichi Endo, Hidetoshi Tanioka, Masaaki Sasaoka, Kei Tashiro, Shigeru Kinoshita, Masaaki Kageyama
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2020
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/592188d10cff4b4fb91119369b563593
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spelling	oai:doaj.org-article:592188d10cff4b4fb91119369b5635932021-12-02T13:34:00ZNovel use of a chemically modified siRNA for robust and sustainable in vivo gene silencing in the retina10.1038/s41598-020-79242-w2045-2322https://doaj.org/article/592188d10cff4b4fb91119369b5635932020-12-01T00:00:00Zhttps://doi.org/10.1038/s41598-020-79242-whttps://doaj.org/toc/2045-2322Abstract Despite efficient and specific in vitro knockdown, more reliable and convenient methods for in vivo knockdown of target genes remain to be developed particularly for retinal research. Using commercially available and chemically modified siRNA so-called Accell siRNA, we established a novel in vivo gene silencing approach in the rat retina. siRNA designed for knockdown of the house keeping gene Gapdh or four retinal cell type-specific genes (Nefl, Pvalb, Rho and Opn1sw) was injected into the vitreous body, and their retinal mRNA levels were quantified using real-time PCR. Intravitreal injection of siRNA for Gapdh resulted in approximately 40–70% reduction in its retinal mRNA levels, which lasted throughout a 9-day study period. Furthermore, all the selected retinal specific genes were efficiently down-regulated by 60–90% following intravitreal injection, suggesting injected siRNA penetrated into major retinal cell types. These findings were consistent with uniform distribution of a fluorescence-labeled siRNA injected into the vitreous body. Interestingly, gene silencing of Grin1, a core subunit of NMDA receptor, was accompanied by significant prevention from NMDA-induced retinal ganglion cell death. Thus, we provide single intravitreal injection of Accell siRNA as a versatile technique for robust and sustainable in vivo retinal gene silencing to characterize their biological functions under physiological and pathophysiological conditions.Takazumi TaniguchiKen-ichi EndoHidetoshi TaniokaMasaaki SasaokaKei TashiroShigeru KinoshitaMasaaki KageyamaNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 10, Iss 1, Pp 1-9 (2020)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Takazumi Taniguchi Ken-ichi Endo Hidetoshi Tanioka Masaaki Sasaoka Kei Tashiro Shigeru Kinoshita Masaaki Kageyama Novel use of a chemically modified siRNA for robust and sustainable in vivo gene silencing in the retina
description	Abstract Despite efficient and specific in vitro knockdown, more reliable and convenient methods for in vivo knockdown of target genes remain to be developed particularly for retinal research. Using commercially available and chemically modified siRNA so-called Accell siRNA, we established a novel in vivo gene silencing approach in the rat retina. siRNA designed for knockdown of the house keeping gene Gapdh or four retinal cell type-specific genes (Nefl, Pvalb, Rho and Opn1sw) was injected into the vitreous body, and their retinal mRNA levels were quantified using real-time PCR. Intravitreal injection of siRNA for Gapdh resulted in approximately 40–70% reduction in its retinal mRNA levels, which lasted throughout a 9-day study period. Furthermore, all the selected retinal specific genes were efficiently down-regulated by 60–90% following intravitreal injection, suggesting injected siRNA penetrated into major retinal cell types. These findings were consistent with uniform distribution of a fluorescence-labeled siRNA injected into the vitreous body. Interestingly, gene silencing of Grin1, a core subunit of NMDA receptor, was accompanied by significant prevention from NMDA-induced retinal ganglion cell death. Thus, we provide single intravitreal injection of Accell siRNA as a versatile technique for robust and sustainable in vivo retinal gene silencing to characterize their biological functions under physiological and pathophysiological conditions.
format	article
author	Takazumi Taniguchi Ken-ichi Endo Hidetoshi Tanioka Masaaki Sasaoka Kei Tashiro Shigeru Kinoshita Masaaki Kageyama
author_facet	Takazumi Taniguchi Ken-ichi Endo Hidetoshi Tanioka Masaaki Sasaoka Kei Tashiro Shigeru Kinoshita Masaaki Kageyama
author_sort	Takazumi Taniguchi
title	Novel use of a chemically modified siRNA for robust and sustainable in vivo gene silencing in the retina
title_short	Novel use of a chemically modified siRNA for robust and sustainable in vivo gene silencing in the retina
title_full	Novel use of a chemically modified siRNA for robust and sustainable in vivo gene silencing in the retina
title_fullStr	Novel use of a chemically modified siRNA for robust and sustainable in vivo gene silencing in the retina
title_full_unstemmed	Novel use of a chemically modified siRNA for robust and sustainable in vivo gene silencing in the retina
title_sort	novel use of a chemically modified sirna for robust and sustainable in vivo gene silencing in the retina
publisher	Nature Portfolio
publishDate	2020
url	https://doaj.org/article/592188d10cff4b4fb91119369b563593
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Novel use of a chemically modified siRNA for robust and sustainable in vivo gene silencing in the retina

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